Osteoarthritis (OA) yog ib lub sij hawm ntev degenerative pob txha cov kab mob uas cuam tshuam rau cov neeg laus dua 65 xyoo.
1]. Feem ntau, OA cov neeg mob tau kuaj pom tias muaj pob txha mos, mob synovium, thiab chondrocytes eroded, uas ua rau mob thiab mob lub cev [
2]. Mob caj dab feem ntau yog tshwm sim los ntawm degeneration ntawm pob txha mos nyob rau hauv cov pob qij txha los ntawm o, thiab thaum cov pob txha mos raug mob hnyav cov pob txha tuaj yeem sib tsoo nrog ib leeg ua rau mob tsis taus thiab mob lub cev [
3]. Kev koom tes ntawm inflammatory mediators nrog cov tsos mob xws li mob, o, thiab txhav ntawm lub pob qij txha yog cov ntaub ntawv zoo. Hauv cov neeg mob OA, inflammatory cytokines, uas ua rau kev yaig ntawm pob txha mos thiab cov pob txha subchondral muaj nyob hauv cov kua dej synovial [
4]. Ob qhov kev tsis txaus siab loj uas cov neeg mob OA feem ntau muaj yog mob thiab mob synovial. Yog li lub hom phiaj tseem ceeb ntawm kev kho OA tam sim no yog txo qhov mob thiab mob. [
5]. Txawm hais tias muaj cov kev kho mob OA, suav nrog cov tshuaj uas tsis yog steroidal thiab steroidal, tau ua pov thawj tias muaj txiaj ntsig zoo hauv kev txo qhov mob thiab mob, kev siv cov tshuaj no mus ntev muaj qhov cuam tshuam rau kev noj qab haus huv xws li mob plawv, plab hnyuv, thiab lub raum tsis ua haujlwm [
6]. Yog li, cov tshuaj muaj txiaj ntsig zoo dua nrog cov kev mob tshwm sim tsawg dua yuav tsum tau tsim rau kev kho mob osteoarthritis.
Cov khoom noj qab haus huv tau nrov zuj zus rau kev nyab xeeb thiab yooj yim muaj [
7]. Cov tshuaj Kaus Lim Kauslim tau ua pov thawj tias muaj txiaj ntsig tiv thaiv ntau yam kab mob, suav nrog kev mob caj dab [
8]. Aucklandia lappa DC. paub txog nws cov khoom siv tshuaj, xws li txhim kho txoj kev ncig ntawm qi rau txo qhov mob thiab soothing lub plab, thiab tau siv ib txwm siv raws li ib qho kev ua kom mob plab [
9]. Cov ntaub ntawv dhau los qhia tias A. lappa muaj cov tshuaj tiv thaiv kab mob [
10,
11], analgesic [
12], anticancer [
13], thiab gastroprotective [
14] teebmeem. Ntau yam kev ua ub no ntawm A. lappa yog tshwm sim los ntawm nws cov ntsiab lus tseem ceeb: costunolide, dehydrocostus lactone, dihydrocostunolide, costuslactone, α-costol, saussurea lactone thiab costuslactone [
15]. Cov kev tshawb fawb yav dhau los tau lees tias costunolide tau pom cov khoom tiv thaiv kab mob hauv lipopolysaccharide (LPS), uas ua rau cov macrophages los ntawm kev tswj hwm ntawm NF-kB thiab cov cua sov ua kom muaj protein ntau txoj hauv kev [
16,
17]. Txawm li cas los xij, tsis muaj kev tshawb fawb tau tshawb xyuas cov dej num muaj peev xwm ntawm A. lappa rau kev kho OA. Cov kev tshawb fawb tam sim no tau tshawb xyuas cov kev kho mob ntawm A. lappa tiv thaiv OA siv (monosodium-iodoacetate) MIA thiab acetic acid-induced nas qauv.
Monosodium-iodoacetate (MIA) yog nto moo siv los tsim ntau yam mob ntawm tus cwj pwm thiab cov yam ntxwv pathophysiological ntawm OA hauv cov tsiaj [
18,
19,
20]. Thaum txhaj rau hauv lub hauv caug pob qij txha, MIA disarrays chondrocyte metabolism thiab induces o thiab inflammatory cov tsos mob, xws li pob txha mos thiab subchondral pob txha erosion, lub cardinal cov tsos mob ntawm OA [
18]. Writhing teb induced nrog acetic acid yog dav suav hais tias yog qhov simulation ntawm peripheral mob nyob rau hauv cov tsiaj uas qhov mob inflammatory yuav ntsuas ntau npaum li cas [
19]. Tus nas macrophage cell kab, RAW264.7, yog nrov siv los kawm cov cellular teb rau o. Thaum ua haujlwm nrog LPS, RAW264 macrophages qhib txoj hauv kev inflammatory thiab zais ntau yam kab mob sib kis, xws li TNF-α, COX-2, IL-1β, iNOS, thiab IL-6 [
20]. Txoj kev tshawb no tau soj ntsuam cov tshuaj tiv thaiv nociceptive thiab anti-inflammatory los ntawm A. lappa tiv thaiv OA hauv MIA tsiaj qauv, acetic acid-induced tsiaj qauv, thiab LPS-activated RAW264.7 hlwb.
2. Cov ntaub ntawv thiab cov txheej txheem
2.1. Khoom siv cog
Cov cag qhuav ntawm A. lappa DC. siv nyob rau hauv qhov kev sim yog procured los ntawm Epulip Pharmaceutical Co., Ltd., (Seoul, Kauslim). Nws raug txheeb xyuas los ntawm Prof. Donghun Lee, Dept. of Herbal pharmacology, Col. ntawm Kauslim Tshuaj, Gachon University, thiab daim ntawv pov thawj tus naj npawb tau muab tso rau hauv 18060301.
2.2. HPLC Kev Ntsuam Xyuas ntawm A. lappa Extract
A. lappa tau muab rho tawm siv lub tshuab reflux (dej distilled, 3 h ntawm 100 ° C). Cov tshuaj tau muab rho tawm tau lim thiab condensed siv lub evaporator uas tsis tshua muaj siab. A. lappa extract muaj cov txiaj ntsig ntawm 44.69% tom qab khov-qhuav hauv qab -80 ° C. Chromatographic tsom xam ntawm A. lappa tau ua nrog HPLC txuas nrog siv 1260 InfinityⅡ HPLC-system (Agilent, Pal Alto, CA, USA). Rau kev sib cais chromatic, EclipseXDB C18 kem (4.6 × 250 mm, 5 µm, Agilent) tau siv ntawm 35 ° C. Tag nrho ntawm 100 mg ntawm cov hnoos qeev tau diluted hauv 10 mL ntawm 50% methanol thiab sonicated rau 10 min. Cov qauv raug lim nrog lub syringe lim (Waters Corp., Milford, MA, USA) ntawm 0.45 μm. Cov theem mobile muaj pes tsawg leeg yog 0.1% phosphoric acid (A) thiab acetonitrile (B) thiab kem tau eluted raws li nram no: 0-60 min, 0%; 60-65 feeb, 100%; 65–67 feeb, 100%; 67-72 min, 0% hnyav B nrog tus nqi ntws ntawm 1.0 mL / min. Cov effluent tau pom ntawm 210 nm siv qhov ntim ntawm 10 μL. Kev tsom xam tau ua nyob rau hauv triplicate.
2.3. Tsiaj Tsev thiab Kev Tswj Xyuas
Txiv neej Sprague-Dawley (SD) nas hnub nyoog 5 lub lis piam thiab txiv neej ICR nas hnub nyoog 6 lub lis piam tau yuav los ntawm Samtako Bio Kauslim (Gyeonggi-do, Kauslim). Tsiaj txhu tau khaws cia rau hauv ib chav uas siv qhov kub thiab txias (22 ± 2 ° C) thiab av noo (55 ± 10%) thiab lub teeb / tsaus voj voog ntawm 12/12 h. Cov tsiaj tau paub txog tus mob rau ntau tshaj ib lub lis piam ua ntej qhov kev sim pib. Tsiaj txhu muaj ad libitum khoom noj thiab dej. Cov kev cai coj ncaj ncees tam sim no rau kev saib xyuas tsiaj thiab tuav ntawm Gachon University (GIACUC-R2019003) tau ua raws li txhua yam kev sim tsiaj. Txoj kev tshawb no tau tsim los ntawm tus kws tshawb nrhiav-dig muag thiab kev sim sib luag. Peb tau ua raws li txoj kev euthanasia raws li cov lus qhia ntawm Pawg Neeg Saib Xyuas Kev Ncaj Ncees Tsiaj.
2.4. MIA Txhaj Tshuaj thiab Kev Kho Mob
Cov nas tsuag tau muab cais ua 4 pawg, xws li sham, tswj, indomethacin, thiab A. lappa. Ua tshuaj loog nrog 2% isofluorane O2 sib tov, cov nas tau txhaj tshuaj siv 50 μL ntawm MIA (40 mg / m; Sigma-Aldrich, St. Louis, MO, USA) intra-articularly rau hauv lub hauv caug pob qij txha los ua rau kev sim OA. Cov kev kho mob tau ua raws li hauv qab no: kev tswj hwm thiab pab pawg sham tsuas yog khaws cia nrog AIN-93G cov zaub mov yooj yim. Tsuas yog, pawg indomethacin tau muab cov tshuaj indomethacin (3 mg / kg) muab tso rau hauv AIN-93G noj zaub mov thiab A. lappa 300 mg / kg pawg tau muab rau AIN-93G noj zaub mov ntxiv nrog A. lappa (300 mg / kg). Cov kev kho mob tau txuas ntxiv rau 24 hnub txij li hnub ntawm OA induction ntawm tus nqi ntawm 15-17 g ib 190-210 g lub cev hnyav rau ib hnub.
2.5. Kev ntsuas qhov hnyav
Tom qab OA induction, qhov hnyav-kev muaj peev xwm ntsuas ntawm hind ceg ntawm cov nas tau ua nrog qhov tsis muaj peev xwm-MeterTester600 (IITC Life Science, Woodland Hills, CA, USA) raws li tau teem tseg. Kev faib tawm qhov hnyav ntawm sab nraub qaum tau suav nrog: qhov hnyav hnyav (%)
